Aryn A. Alanizi, Alexandre M. Sorlin, Matthew F. L. Parker, Marina López-Álvarez, Hecong Qin, Sang Hee Lee, Joseph Blecha, Oren S. Rosenberg, Joanne Engel, Michael A. Ohliger, Robert R. Flavell and David M. Wilson*,
{"title":"使用 [18F]FB-sulfo-DBCO 对叠氮修饰细菌进行生物正交放射标记。","authors":"Aryn A. Alanizi, Alexandre M. Sorlin, Matthew F. L. Parker, Marina López-Álvarez, Hecong Qin, Sang Hee Lee, Joseph Blecha, Oren S. Rosenberg, Joanne Engel, Michael A. Ohliger, Robert R. Flavell and David M. Wilson*, ","doi":"10.1021/acs.bioconjchem.4c00024","DOIUrl":null,"url":null,"abstract":"<p ><i>Purpose:</i> This study was motivated by the need for better positron emission tomography (PET)-compatible tools to image bacterial infection. Our previous efforts have targeted bacteria-specific metabolism via assimilation of carbon-11 labeled <span>d</span>-amino acids into the bacterial cell wall. Since the chemical determinants of this incorporation are not fully understood, we sought a high-throughput method to label <span>d</span>-amino acid derived structures with fluorine-18. Our strategy employed a chemical biology approach, whereby an azide (-N<sub>3</sub>) bearing <span>d</span>-amino acid is incorporated into peptidoglycan muropeptides, with subsequent “click” cycloaddition with an <sup>18</sup>F-labeled strained cyclooctyne partner. <i>Procedures:</i> A water-soluble, <sup>18</sup>F-labeled and dibenzocyclooctyne (DBCO)-derived radiotracer ([<sup>18</sup>F]FB-sulfo-DBCO) was synthesized. This tracer was incubated with pathogenic bacteria treated with azide-bearing <span>d</span>-amino acids, and incorporated <sup>18</sup>F was determined via gamma counting. <i>In vitro</i> uptake in bacteria previously treated with azide-modified <span>d</span>-amino acids was compared to that in cultures treated with amino acid controls. The biodistribution of [<sup>18</sup>F]FB-sulfo-DBCO was studied in a cohort of healthy mice with implications for future <i>in vivo</i> imaging. <i>Results:</i> The new strain-promoted azide–alkyne cycloaddition (SPAAC) radiotracer [<sup>18</sup>F]FB-sulfo-DBCO was synthesized with high radiochemical yield and purity via <i>N</i>-succinimidyl 4-[<sup>18</sup>F]fluorobenzoate ([<sup>18</sup>F]SFB). Accumulation of [<sup>18</sup>F]FB-sulfo-DBCO was significantly higher in several bacteria treated with azide-modified <span>d</span>-amino acids than in controls; for example, we observed 7 times greater [<sup>18</sup>F]FB-sulfo-DBCO ligation in <i>Staphylococcus aureus</i> cultures incubated with 3-azido-<span>d</span>-alanine versus those incubated with <span>d</span>-alanine. <i>Conclusions:</i> The SPAAC radiotracer [<sup>18</sup>F]FB-sulfo-DBCO was validated <i>in vitro</i> via metabolic labeling of azide-bearing peptidoglycan muropeptides. <span>d</span>-Amino acid-derived PET radiotracers may be more efficiently screened via [<sup>18</sup>F]FB-sulfo-DBCO modification.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry Bioconjugate","volume":null,"pages":null},"PeriodicalIF":4.0000,"publicationDate":"2024-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.4c00024","citationCount":"0","resultStr":"{\"title\":\"Bioorthogonal Radiolabeling of Azide-Modified Bacteria Using [18F]FB-sulfo-DBCO\",\"authors\":\"Aryn A. Alanizi, Alexandre M. Sorlin, Matthew F. L. Parker, Marina López-Álvarez, Hecong Qin, Sang Hee Lee, Joseph Blecha, Oren S. Rosenberg, Joanne Engel, Michael A. Ohliger, Robert R. Flavell and David M. Wilson*, \",\"doi\":\"10.1021/acs.bioconjchem.4c00024\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p ><i>Purpose:</i> This study was motivated by the need for better positron emission tomography (PET)-compatible tools to image bacterial infection. Our previous efforts have targeted bacteria-specific metabolism via assimilation of carbon-11 labeled <span>d</span>-amino acids into the bacterial cell wall. Since the chemical determinants of this incorporation are not fully understood, we sought a high-throughput method to label <span>d</span>-amino acid derived structures with fluorine-18. Our strategy employed a chemical biology approach, whereby an azide (-N<sub>3</sub>) bearing <span>d</span>-amino acid is incorporated into peptidoglycan muropeptides, with subsequent “click” cycloaddition with an <sup>18</sup>F-labeled strained cyclooctyne partner. <i>Procedures:</i> A water-soluble, <sup>18</sup>F-labeled and dibenzocyclooctyne (DBCO)-derived radiotracer ([<sup>18</sup>F]FB-sulfo-DBCO) was synthesized. This tracer was incubated with pathogenic bacteria treated with azide-bearing <span>d</span>-amino acids, and incorporated <sup>18</sup>F was determined via gamma counting. <i>In vitro</i> uptake in bacteria previously treated with azide-modified <span>d</span>-amino acids was compared to that in cultures treated with amino acid controls. The biodistribution of [<sup>18</sup>F]FB-sulfo-DBCO was studied in a cohort of healthy mice with implications for future <i>in vivo</i> imaging. <i>Results:</i> The new strain-promoted azide–alkyne cycloaddition (SPAAC) radiotracer [<sup>18</sup>F]FB-sulfo-DBCO was synthesized with high radiochemical yield and purity via <i>N</i>-succinimidyl 4-[<sup>18</sup>F]fluorobenzoate ([<sup>18</sup>F]SFB). Accumulation of [<sup>18</sup>F]FB-sulfo-DBCO was significantly higher in several bacteria treated with azide-modified <span>d</span>-amino acids than in controls; for example, we observed 7 times greater [<sup>18</sup>F]FB-sulfo-DBCO ligation in <i>Staphylococcus aureus</i> cultures incubated with 3-azido-<span>d</span>-alanine versus those incubated with <span>d</span>-alanine. <i>Conclusions:</i> The SPAAC radiotracer [<sup>18</sup>F]FB-sulfo-DBCO was validated <i>in vitro</i> via metabolic labeling of azide-bearing peptidoglycan muropeptides. <span>d</span>-Amino acid-derived PET radiotracers may be more efficiently screened via [<sup>18</sup>F]FB-sulfo-DBCO modification.</p>\",\"PeriodicalId\":29,\"journal\":{\"name\":\"Bioconjugate Chemistry Bioconjugate\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.0000,\"publicationDate\":\"2024-03-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://pubs.acs.org/doi/epdf/10.1021/acs.bioconjchem.4c00024\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bioconjugate Chemistry Bioconjugate\",\"FirstCategoryId\":\"1\",\"ListUrlMain\":\"https://pubs.acs.org/doi/10.1021/acs.bioconjchem.4c00024\",\"RegionNum\":2,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioconjugate Chemistry Bioconjugate","FirstCategoryId":"1","ListUrlMain":"https://pubs.acs.org/doi/10.1021/acs.bioconjchem.4c00024","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Bioorthogonal Radiolabeling of Azide-Modified Bacteria Using [18F]FB-sulfo-DBCO
Purpose: This study was motivated by the need for better positron emission tomography (PET)-compatible tools to image bacterial infection. Our previous efforts have targeted bacteria-specific metabolism via assimilation of carbon-11 labeled d-amino acids into the bacterial cell wall. Since the chemical determinants of this incorporation are not fully understood, we sought a high-throughput method to label d-amino acid derived structures with fluorine-18. Our strategy employed a chemical biology approach, whereby an azide (-N3) bearing d-amino acid is incorporated into peptidoglycan muropeptides, with subsequent “click” cycloaddition with an 18F-labeled strained cyclooctyne partner. Procedures: A water-soluble, 18F-labeled and dibenzocyclooctyne (DBCO)-derived radiotracer ([18F]FB-sulfo-DBCO) was synthesized. This tracer was incubated with pathogenic bacteria treated with azide-bearing d-amino acids, and incorporated 18F was determined via gamma counting. In vitro uptake in bacteria previously treated with azide-modified d-amino acids was compared to that in cultures treated with amino acid controls. The biodistribution of [18F]FB-sulfo-DBCO was studied in a cohort of healthy mice with implications for future in vivo imaging. Results: The new strain-promoted azide–alkyne cycloaddition (SPAAC) radiotracer [18F]FB-sulfo-DBCO was synthesized with high radiochemical yield and purity via N-succinimidyl 4-[18F]fluorobenzoate ([18F]SFB). Accumulation of [18F]FB-sulfo-DBCO was significantly higher in several bacteria treated with azide-modified d-amino acids than in controls; for example, we observed 7 times greater [18F]FB-sulfo-DBCO ligation in Staphylococcus aureus cultures incubated with 3-azido-d-alanine versus those incubated with d-alanine. Conclusions: The SPAAC radiotracer [18F]FB-sulfo-DBCO was validated in vitro via metabolic labeling of azide-bearing peptidoglycan muropeptides. d-Amino acid-derived PET radiotracers may be more efficiently screened via [18F]FB-sulfo-DBCO modification.
期刊介绍:
Bioconjugate Chemistry invites original contributions on all research at the interface between man-made and biological materials. The mission of the journal is to communicate to advances in fields including therapeutic delivery, imaging, bionanotechnology, and synthetic biology. Bioconjugate Chemistry is intended to provide a forum for presentation of research relevant to all aspects of bioconjugates, including the preparation, properties and applications of biomolecular conjugates.